摘要
目的探讨miR-146a对PC-12细胞帕金森病(PD)模型的生物学行为的影响,及其介导NLRP3对细胞模型焦亡机制。方法各采集16例PD患者、健康人群外周血,实时荧光定量PCR检测miR-146a在PD患者外周血和正常人群外周血中的表达量。采用LPS干预PC-12细胞,建立PD模型。将miR-146a模拟物、抑制剂分别转染PC-12 PD模型细胞,采用CCK-8检测细胞增殖活性变化,探讨miR-146a对细胞表型的调控作用。采用ELISA检测白细胞介素IL-1α(IL-1β)、IL-18和LDH的表达变化,实时荧光定量PCR检测肿瘤坏死因子α(TNF-α)的mRNA表达差异性,蛋白印迹(Western blot)检测pro-caspase-1、caspase-1、NLRP3的蛋白表达差异性,探讨miR-146a对PC-12细胞焦亡的调控机制作用。结果miR-146a在PD患者外周血中呈低表达。miR-146a mimics会抑制PC-12细胞增殖活性,提高IL-1β、IL-18、LDH、TNF-α、caspase-1、NLRP3的表达(P<0.05)。miR-146a inhibitor会促进PC-12细胞增殖活性,抑制IL-1β、IL-18、乳酸脱氢酶(LDH)、TNF-α、caspase-1、NLRP3的表达(P<0.05)。结论miR-146a可以起到抑制细胞增殖活性,诱导细胞凋亡,加剧炎性因子和焦亡相关蛋白表达,诱导PD的发生发展。
Objective Exploring the biological behavior of PC-12 cells in Parkinson's disease(PD)model and the mediating mechanism of NLRP3 induced apoptosis in the cell model by miR-146a.Methods Collect peripheral blood samples from 16 PD patients and healthy individuals,and use real-time fluorescence quantitative PCR to detect the expression levels of miR-146a in the peripheral blood of PD patients and normal individuals.Using LPS intervention on PC-12 cells to establish a PD model.Transfect PC-12 PD model cells with miR-146a mimetics and inhibitors,and detect changes in cell proliferation activity using CCK-8 to investigate the regulatory effect of miR-146a on cell phenotype.ELISA was used to detect the expression changes of IL-1β,IL-18,and LDH,real-time fluorescence quantitative PCR was used to detect the differential mRNA expression of TNF-α,and Western blot was used to detect the differential protein expression of pro-caspase-1,caspase-1,and NLRP3.The regulatory mechanism of miR-146a on apoptosis of PC-12 cells was explored.Results miR-146a is lowly expressed in the peripheral blood of PD patients.miR-146a mimetics can inhibit the proliferation activity of PC-12 cells and increase the expression of IL-1β,IL-18,LDH,TNF-α,caspase-1,and NLRP3(P<0.05).miR-146a inhibitor promotes the proliferation activity of PC-12 cells and inhibits the expression of IL-1β,IL-18,LDH,TNF-α,caspase-1,and NLRP3 P<0.05.Conclusion miR-146a can inhibit cell proliferation,induce cell apoptosis,and increase the expression of inflammatory PD.
作者
陶静
赵洋
李金贤
Tao Jing;Zhao Yang;Li Jinxian(Rehabilitation Department of Xinjiang Uygur Autonomous Region People's Hospital,Urumqi 830000,China)
出处
《脑与神经疾病杂志》
2025年第5期277-282,共6页
Journal of Brain and Nervous Diseases
基金
新疆维吾尔自治区自然科学基金项目(2022D01C125)
新疆维吾尔自治区人民医院院内项目(20190414)。
